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Heterologous Expression of Membrane Proteins pp 49–66Cite as

Expression of Membrane Proteins at the Escherichia coli Membrane for Structural Studies

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 601))

Abstract

Structural biology of membrane proteins is often limited by the first steps in obtaining sufficient yields of proteins because native sources are seldom. Heterologous systems like bacteria are then commonly employed for membrane protein over-expression. Escherichia coli is the main bacterial host used. However, overproduction of a foreign membrane protein at a non-physiological level is usually toxic for cells or leads to inclusion body formation. Those effects can be reduced by optimizing the cell growth conditions, choosing the suitable bacterial strain and expression vector, and finally co-expressing the target protein and the b-subunit of E. coli adenosine triphosphate (ATP)-synthase, which triggers the proliferation of intracytoplasmic membranes. This chapter is devoted to help the experimenter in choosing the appropriate plasmid/bacterial host combination for optimizing the amount of the target membrane protein produced in its correct folded state.

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Acknowledgments

This work was supported by INSERM, CNRS and the “Agence Nationale de la Recherche” (grant 05-JCJC-0092-01 to B.M.).

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Authors and Affiliations

  1. Université Paris-7, Laboratoire de Biologie Physico-Chimique des Proteines Membranaires, Institut de Biologie Physico-Chimique, Paris, France

    Manuela Zoonens & Bruno Miroux

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  1. Manuela Zoonens
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  2. Bruno Miroux
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Corresponding author

Correspondence to Bruno Miroux .

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Editors and Affiliations

  1. CNRS - UMR 6543, Université Nice, parc Valrose, Nice CX 02, 06108, France

    Isabelle Mus-Veteau

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Zoonens, M., Miroux, B. (2010). Expression of Membrane Proteins at the Escherichia coli Membrane for Structural Studies. In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology™, vol 601. Humana Press. https://doi.org/10.1007/978-1-60761-344-2_4

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  • DOI: https://doi.org/10.1007/978-1-60761-344-2_4

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-343-5

  • Online ISBN: 978-1-60761-344-2

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